RADseq Library Preparation Protocol

RADseq libraries were prepared from the isolated DNA as described following Grewe et al. (2017 (link)). In summary, DNA isolations were pooled with sequence adapters (Rubin & Moreau, 2016 (link)), digested with the restriction enzyme ApeKI (New England Biolabs, Ipswich, MA, USA), and ligated using T4 ligase (New England Biolabs). All samples with compatible barcodes were pooled and selected for fragment sizes between 300 and 500 bp using the BluePippin DNA size selection system (Sage Science, Beverly, MA, USA). The pooled libraries were amplified using the REDTaq ReadyMix (Sigma‐Aldrich, St. Louis, MO, USA) prior to sequencing on an Illumina MiSeq using the MiSeq Reagent Kit v3 for 150 cycles (Illumina, San Diego, CA, USA) to produce single‐end sequences with a length of 150 bp.